Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F13%3A00072093" target="_blank" >RIV/00216224:14740/13:00072093 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/68081715:_____/13:00398549

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.chroma.2013.10.087" target="_blank" >http://dx.doi.org/10.1016/j.chroma.2013.10.087</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.chroma.2013.10.087" target="_blank" >10.1016/j.chroma.2013.10.087</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis

Popis výsledku v původním jazyce

In this paper, we report on a novel oriented peptide-N-glycosidase F (PNGase F) immobilization approach onto methacrylate based monolithic support for rapid, reproducible and efficient release of the N-linked carbohydrate moieties from glycoproteins. Theglutathione-S-transferase-fusion PNGase F (PNGase F-GST) was expressed in Escherichia coli using regular vector technology. The monolithic pore surface was functionalized with glutathione via a succinimidy1-6-(iodoacetyl-amino)-hexanoate linker and thespecific affinity of GST toward glutathione was utilized for the oriented coupling. This novel immobilization procedure was compared with reductive amination technique commonly used for non-oriented enzyme immobilization via primary amine functionalities. Both coupling approaches were compared using enzymatic treatment of several glycoproteins, such as ribonuclease B, fetuin and immunoglobulin G followed by MALDI/MS and CE-LIF analysis of the released glycans.

Název v anglickém jazyce

Oriented immobilization of peptide-N-glycosidase F on a monolithic support for glycosylation analysis

Popis výsledku anglicky

In this paper, we report on a novel oriented peptide-N-glycosidase F (PNGase F) immobilization approach onto methacrylate based monolithic support for rapid, reproducible and efficient release of the N-linked carbohydrate moieties from glycoproteins. Theglutathione-S-transferase-fusion PNGase F (PNGase F-GST) was expressed in Escherichia coli using regular vector technology. The monolithic pore surface was functionalized with glutathione via a succinimidy1-6-(iodoacetyl-amino)-hexanoate linker and thespecific affinity of GST toward glutathione was utilized for the oriented coupling. This novel immobilization procedure was compared with reductive amination technique commonly used for non-oriented enzyme immobilization via primary amine functionalities. Both coupling approaches were compared using enzymatic treatment of several glycoproteins, such as ribonuclease B, fetuin and immunoglobulin G followed by MALDI/MS and CE-LIF analysis of the released glycans.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CB - Analytická chemie, separace

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2013

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Chromatography A

ISSN

0021-9673

e-ISSN

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Svazek periodika

1322

Číslo periodika v rámci svazku

Dec

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

8

Strana od-do

54-61

Kód UT WoS článku

000328717200007

EID výsledku v databázi Scopus

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